LOCUS       sequence-422953-        6200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This binary vector expresses mushroom luciferase (nnLuz) containing 
            the PIV intron (216 bp into the gene) with the truncated 35sCAMV 
            promoter and Tomato mosaic virus UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6200)
  TITLE     Curtis lab plasmids
REFERENCE   2  (bases 1 to 6200)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLSU1/t35s[TMV]:nnLuz-I{PIV-216}:NosT
FEATURES             Location/Qualifiers
     source          1..6200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             723..1796
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      1862..2056
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     2519..2539
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        2528..2574
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     intron          2894..3082
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(3337..3348)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      3695..3947
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4123..4142)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4134..4949
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4134..4949
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4207..4226)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5035..5623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5524..5543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5865..6200,1..294)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
       61 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
      121 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
      181 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
      241 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
      301 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
      361 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
      421 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
      481 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
      541 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
      601 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
      661 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
      721 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
      781 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
      841 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
      901 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
      961 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     1021 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     1081 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     1141 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     1201 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     1261 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     1321 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     1381 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     1441 acatcgagat cgagctagtt gattggatgt accgcgagat cacagaaggc aagaacccgg
     1501 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     1561 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     1621 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     1681 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     1741 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     1801 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa gatctctttc
     1861 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     1921 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     1981 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     2041 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
     2101 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
     2161 gccgctcaaa aatggctggc ctacggccag gcaatctgct agcgcgcgga caagctagga
     2221 ttggcaggat atattcagtt gtaaatgata tcgcatgcgg aaacctcctc ggattccatt
     2281 gcccagctat ctgtcacttt attgagaaga tagtggaaaa ggaaggtggc tcctacaaat
     2341 gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca
     2401 aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt
     2461 caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact
     2521 atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag aggacacgct
     2581 cgagtataag agctcatttt tacaacaatt accaacaaca acaaacaaca aacaacatta
     2641 caattacatt tacaattatc gatacatcga gggtaccatg aggataaata tctctttgtc
     2701 tagtctcttt gagagactga gcaaattgtc atcccgatcc atcgcaatta cctgcggcgt
     2761 agtcttggca agtgcaatag cattcccaat tatccgaaga gactatcaaa cgttccttga
     2821 agtcggtccg agctatgccc cacagaactt ccgaggctat atcatcgttt gtgtcttgtc
     2881 actttttagg caagtaagtt tctgcttcta cctttgatat atatataata attatcatta
     2941 attagtagta atataatatt tcaaatattt ttttcaaaat aaaagaatgt agtatatagc
     3001 aattgctttt ctgtagttta taagtgtgta tattttaatt tataactttt ctaatatatg
     3061 accaaaattt gttgatgtgc aggagcagaa agggttggca atctatgaca ggcttccaga
     3121 aaagaggcgt tggcttgccg acttgccgtt tcgtgaaggg acgaggccat ccataacctc
     3181 ccacattata cagcgtcagc gtactcagct ggtagatcaa gagtttgcaa ccagagaact
     3241 tatcgataag gtgatcccac gtgtgcaagc aagacacaca gacaaaactt ttctcagtac
     3301 gtcaaaattt gagtttcatg caaaggccat cttccttctc ccctctatcc ctattaatga
     3361 tcctcttaac attccctcac acgatacggt aagaagaacc aagcgtgaga tcgctcacat
     3421 gcatgattac catgattgca ctctgcactt ggctcttgct gctcaggatg gtaaggaagt
     3481 tttgaagaag gggtggggcc agcgtcaccc actggccgga ccaggagtcc caggccctcc
     3541 tactgagtgg accttccttt acgcaccaag gaacgaagag gaggccagag ttgtcgagat
     3601 gatagtcgaa gctagtattg gctacatgac taacgatcct gctggtaaga ttgtcgaaaa
     3661 tgccaagtca ggttgagaat tcggatccct gcaggatcgt tcaaacattt ggcaataaag
     3721 tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa
     3781 ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt
     3841 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc
     3901 aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg gagctctggc
     3961 aggatatatg cggttgtaat tcatttttat tgtctaaatt tctgtatttg tttgtttgca
     4021 attagatcca cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4081 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     4141 atattcaacg ggaaacgtct tgttcgaggc cgcgattaaa ttccaacatg gatgctgatt
     4201 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctaccgat
     4261 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     4321 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     4381 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatcccag
     4441 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     4501 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     4561 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     4621 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     4681 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     4741 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     4801 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     4861 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     4921 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     4981 agagcattac gctgacttga cgggacggcg gctttgttga atcaattgat cttcttgaaa
     5041 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5101 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5161 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5221 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5281 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5341 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5401 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5461 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5521 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5581 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5641 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5701 ccctgattct gtggataacc gtattaccgc ctttgagtga tctctgaaga aaccgagcgc
     5761 cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg
     5821 tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg
     5881 tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc
     5941 tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg
     6001 attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga
     6061 ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc
     6121 aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc
     6181 agccaagccc ttacgacata
//
